Part I: development of novel fluorescence quenchers, part II: improving catalysis of hydrazone and oxime formation at biological pH
- This thesis is composed of two parts. In the first part, the development of novel fluorescence quenchers is discussed. The design and properties of Multiple Pathway Quenchers (MPQs) that display broadened absorption properties that allow for greater quenching of a wider variety of fluorophores than common commercial quenchers is examined. The prepared quenchers display complex electronic pathways that provide considerably red-shifted absorption maxima and widened spectra relative to the parent dyes. Particularly in FRET quenching, these new compounds are observed to perform better than other quenchers previously reported. Further studies discuss new quenchers designed for oxime and hydrazone bioorthogonal attachment to fluorescently labeled biomolecules. Hydrazine-containing dyes in particular afford powerful properties for fluorescence quenchers upon hydrazone formation. These quenchers allow for new means of attaching fluorescent molecules to biologically relevant probes and provide the first example of quenching a fluorophore in a living cell without the need for photobleaching. The second part of this thesis explores the design and properties of new catalysts for bioorthogonal hydrazone and oxime formation at physiological pH. Traditionally, aniline has served as the recommended catalyst for hydrazone or oxime formation at pH 7.4, but exceedingly high concentrations are required that hinder simpler applications. Anthranilic acid derivatives are reported as the first enhanced catalysis for both hydrazone and oxime formation at pH 7.4. Further studies display that optimizing the pKa of the ortho proton donor in the anthranilic acid scaffold to match that of the buffer pH affords further enhanced catalysis, with 2-aminobenzenephosphonic acids discussed as the optimal class of catalysts at biological pH.
|̀Improving catalysis of hydrazone and oxime formation at biological pH
|Type of resource
|electronic; electronic resource; remote
|1 online resource.
|Crisalli, Peter Joseph
|Stanford University, Department of Chemistry.
|Kool, Eric T
|Kool, Eric T
|Statement of responsibility
|Peter Joseph Crisalli.
|Submitted to the Department of Chemistry.
|Thesis (Ph.D.)--Stanford University, 2013.
- © 2013 by Peter Joseph Crisalli
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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